{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T18:59:04Z","timestamp":1785524344301,"version":"3.56.0"},"reference-count":42,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T00:00:00Z","timestamp":1760486400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004663","name":"the National Key R&D Program","doi-asserted-by":"publisher","award":["2022YFD1300301"],"award-info":[{"award-number":["2022YFD1300301"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004663","name":"Ningbo Major Science and Technology Project","doi-asserted-by":"publisher","award":["2021Z112"],"award-info":[{"award-number":["2021Z112"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004663","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31930103"],"award-info":[{"award-number":["31930103"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>C-type natriuretic peptide (CNP), encoded by the NPPC (Natriuretic Peptide Precursor C), has been recognized as the principal endogenous factor sustaining oocyte meiotic arrest in mammalian follicles. Yet its influence on porcine ovarian granulosa cell fate and the regulatory mechanism of NPPC expression within these cells remain poorly understood. Here, utilizing an in vitro culture model of primary porcine ovarian granulosa cells and immature oocytes, we examined the impact of CNP on granulosa cell apoptosis and oocyte meiotic resumption, and elucidated the molecular circuitry governing NPPC expression. We found that follicular atresia in pigs was accompanied by a marked decline in the CNP receptor NPR2 (natriuretic peptide receptor 2). Correspondingly, exogenous CNP suppressed apoptosis in cultured porcine granulosa cells. Estradiol can significantly promote the expression level of NPPC in porcine ovarian granulosa cells and, by enhancing NPR2 levels, it can synergize with CNP to inhibit oocyte meiotic resumption in vitro. Conversely, EGF signaling can significantly downregulate NPPC mRNA expression in porcine granulosa cells, an effect likely mediated by ERK-activated tristetraprolin (TTP). Collectively, these findings broaden our understanding of CNP in follicular development and delineate the endocrine network that controls NPPC transcription in the porcine ovary.<\/jats:p>","DOI":"10.3390\/ijms262010046","type":"journal-article","created":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T14:04:02Z","timestamp":1760537042000},"page":"10046","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["The Anti-Apoptotic Effect of C-Type Natriuretic Peptide and the Regulation of NPPC in Porcine Ovarian Granulosa Cells"],"prefix":"10.3390","volume":"26","author":[{"given":"Xingyuan","family":"Liu","sequence":"first","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture and Rural Affairs, Frontiers Science Center for Molecular Design Breeding (MOE), College of Animal Science and Technology, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinlun","family":"Lu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture and Rural Affairs, Frontiers Science Center for Molecular Design Breeding (MOE), College of Animal Science and Technology, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-2137-7482","authenticated-orcid":false,"given":"Junyi","family":"Zeng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture and Rural Affairs, Frontiers Science Center for Molecular Design Breeding (MOE), College of Animal Science and Technology, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"An","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture and Rural Affairs, Frontiers Science Center for Molecular Design Breeding (MOE), College of Animal Science and Technology, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianhui","family":"Tian","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture and Rural Affairs, Frontiers Science Center for Molecular Design Breeding (MOE), College of Animal Science and Technology, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangyin","family":"Xi","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture and Rural Affairs, Frontiers Science Center for Molecular Design Breeding (MOE), College of Animal Science and Technology, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Rintz, E., Wegrzyn, G., Fujii, T., and Tomatsu, S. (2022). Molecular Mechanism of Induction of Bone Growth by the C-Type Natriuretic Peptide. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23115916"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1210\/er.2005-0014","article-title":"Natriuretic peptides, their receptors, and cyclic guanosine monophosphate-dependent signaling functions","volume":"27","author":"Potter","year":"2006","journal-title":"Endocr. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Straczynska, P., Papis, K., Morawiec, E., Czerwinski, M., Gajewski, Z., Olejek, A., and Bednarska-Czerwinska, A. (2022). Signaling mechanisms and their regulation during in vivo or in vitro maturation of mammalian oocytes. Reprod. Biol. Endocrin., 20.","DOI":"10.1186\/s12958-022-00906-5"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.theriogenology.2017.09.003","article-title":"Natriuretic peptide receptor 2 (NPR2) localized in bovine oocyte underlies a unique mechanism for C-type natriuretic peptide (CNP)-induced meiotic arrest","volume":"106","author":"Xi","year":"2018","journal-title":"Theriogenology"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Xu, R., Pan, M., Yin, L., Zhang, Y., Tang, Y., Lu, S., Gao, Y., Wei, Q., Han, B., and Ma, B. (2023). C-Type Natriuretic Peptide Pre-Treatment Improves Maturation Rate of Goat Oocytes by Maintaining Transzonal Projections, Spindle Morphology, and Mitochondrial Function. Animals, 13.","DOI":"10.3390\/ani13243880"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.bbrc.2018.02.054","article-title":"Effects of pre-incubation with C-type natriuretic peptide on nuclear maturation, mitochondrial behavior, and developmental competence of sheep oocytes","volume":"497","author":"Zhang","year":"2018","journal-title":"Biochem. Bioph. Res. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Wang, H., Liu, W., Yang, Y., Wang, X., Zhang, Z., Guo, Q., Wang, C., and Xia, G. (2017). Natriuretic peptides improve the developmental competence of in vitro cultured porcine oocytes. Reprod. Biol. Endocrin., 15.","DOI":"10.1186\/s12958-017-0258-1"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1093\/humupd\/dmad023","article-title":"A fresh start for IVM: Capacitating the oocyte for development using pre-IVM","volume":"30","author":"Gilchrist","year":"2024","journal-title":"Hum. Reprod. Update"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Fakhar-I-Adil, M., Angel-Velez, D., Araftpoor, E., Amin, Q.A., Hedia, M., B\u00fchler, M., Gevaert, K., Menten, B., Van Soom, A., and Chuva De Sousa Lopes, S.M. (2025). Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development Without Inducing Genetic Aberrations. Int. J. Mol. Sci., 26.","DOI":"10.20944\/preprints202505.1132.v1"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1553","DOI":"10.1071\/RD14425","article-title":"C-Type natriuretic peptide maintains domestic cat oocytes in meiotic arrest","volume":"28","author":"Zhong","year":"2016","journal-title":"Reprod. Fert. Develop."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e23295","DOI":"10.1096\/fj.202300886R","article-title":"\u201cMeiosis arrester\u201d C-natriuretic peptide directly stimulates oocyte mtDNA accumulation and is implicated in aging-associated oocyte mtDNA loss","volume":"37","author":"Tang","year":"2023","journal-title":"Faseb J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"RP88523","DOI":"10.7554\/eLife.88523.3","article-title":"C-type natriuretic peptide improves maternally aged oocytes quality by inhibiting excessive PINK1\/Parkin-mediated mitophagy","volume":"12","author":"Zhang","year":"2023","journal-title":"Elife"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4377","DOI":"10.1210\/en.2011-1118","article-title":"Estradiol promotes and maintains cumulus cell expression of natriuretic peptide receptor 2 (NPR2) and meiotic arrest in mouse oocytes in vitro","volume":"152","author":"Zhang","year":"2011","journal-title":"Endocrinology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.1002\/mrd.22424","article-title":"Epidermal growth factor-network signaling mediates luteinizing hormone regulation of BNP and CNP and their receptor NPR2 during porcine oocyte meiotic resumption","volume":"81","author":"Zhang","year":"2014","journal-title":"Mol. Reprod. Dev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3094","DOI":"10.1093\/humrep\/der282","article-title":"Pre-ovulatory LH\/hCG surge decreases C-type natriuretic peptide secretion by ovarian granulosa cells to promote meiotic resumption of pre-ovulatory oocytes","volume":"26","author":"Kawamura","year":"2011","journal-title":"Hum. Reprod."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1038\/s41419-019-1797-5","article-title":"Transforming growth factor-beta is involved in maintaining oocyte meiotic arrest by promoting natriuretic peptide type C expression in mouse granulosa cells","volume":"10","author":"Yang","year":"2019","journal-title":"Cell Death Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1126\/science.1193573","article-title":"Granulosa cell ligand NPPC and its receptor NPR2 maintain meiotic arrest in mouse oocytes","volume":"330","author":"Zhang","year":"2010","journal-title":"Science"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e1475","DOI":"10.1038\/cddis.2014.400","article-title":"Involvement of FoxO1 in the effects of follicle-stimulating hormone on inhibition of apoptosis in mouse granulosa cells","volume":"5","author":"Shen","year":"2014","journal-title":"Cell Death Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1210\/me.2012-1307","article-title":"IGF-I signaling is essential for FSH stimulation of AKT and steroidogenic genes in granulosa cells","volume":"27","author":"Zhou","year":"2013","journal-title":"Mol. Endocrinol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"bqaa038","DOI":"10.1210\/endocr\/bqaa038","article-title":"The FSH-HIF-1alpha-VEGF Pathway Is Critical for Ovulation and Oocyte Health but Not Necessary for Follicular Growth in Mice","volume":"161","author":"Li","year":"2020","journal-title":"Endocrinology"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Casarini, L., Riccetti, L., De Pascali, F., Gilioli, L., Marino, M., Vecchi, E., Morini, D., Nicoli, A., La Sala, G.B., and Simoni, M. (2017). Estrogen Modulates Specific Life and Death Signals Induced by LH and hCG in Human Primary Granulosa Cells In Vitro. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18050926"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Regan, S.L.P., Knight, P.G., Yovich, J.L., Leung, Y., Arfuso, F., and Dharmarajan, A. (2018). Granulosa Cell Apoptosis in the Ovarian Follicle-A Changing View. Front. Endocrinol., 9.","DOI":"10.3389\/fendo.2018.00061"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2204","DOI":"10.1210\/endo.133.5.8404672","article-title":"Estrogens inhibit and androgens enhance ovarian granulosa cell apoptosis","volume":"133","author":"Billig","year":"1993","journal-title":"Endocrinology"},{"key":"ref_24","first-page":"495","article-title":"Mechanisms of atresia in ovarian follicles","volume":"6","author":"Celestino","year":"2009","journal-title":"Anim. Reprod."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Xi, H., Chen, X., Wang, X., Jiang, F., and Niu, D. (2025). Role of programmed cell death in mammalian ovarian follicular atresia. J. Steroid Biochem., 247.","DOI":"10.1016\/j.jsbmb.2024.106667"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Lu, J., Guo, M., Wang, X., Wang, R., Xi, G., An, L., Tian, J., and Chu, M. (2023). A Redesigned Method for CNP-Synchronized In Vitro Maturation Inhibits Oxidative Stress and Apoptosis in Cumulus-Oocyte Complexes and Improves the Developmental Potential of Porcine Oocytes. Genes, 14.","DOI":"10.3390\/genes14101885"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1002\/mrd.22114","article-title":"CNP\/NPR2 signaling maintains oocyte meiotic arrest in early antral follicles and is suppressed by EGFR-mediated signaling in preovulatory follicles","volume":"79","author":"Tsuji","year":"2012","journal-title":"Mol. Reprod. Dev."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e2018345118","DOI":"10.1073\/pnas.2018345118","article-title":"The mRNA-destabilizing protein Tristetraprolin targets \u201cmeiosis arrester\u201d Nppc mRNA in mammalian preovulatory follicles","volume":"118","author":"Xi","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.anireprosci.2004.04.031","article-title":"Follicular development: The role of the follicular microenvironment in selection of the dominant follicle","volume":"82\u201383","author":"Fortune","year":"2004","journal-title":"Anim. Reprod. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1523","DOI":"10.1007\/s11259-023-10107-z","article-title":"The effects of apelin on IGF1\/FSH-induced steroidogenesis, proliferation, Bax expression, and total antioxidant capacity in granulosa cells of buffalo ovarian follicles","volume":"47","author":"Shokrollahi","year":"2023","journal-title":"Vet. Res. Commun."},{"key":"ref_31","first-page":"172","article-title":"Reproductive roles of novel adipokines apelin, visfatin, and irisin in farm animals","volume":"82","author":"Shokrollahi","year":"2021","journal-title":"Theriogenology"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1530\/REP-12-0050","article-title":"NPPC\/NPR2 signaling is essential for oocyte meiotic arrest and cumulus oophorus formation during follicular development in the mouse ovary","volume":"144","author":"Kiyosu","year":"2012","journal-title":"Reproduction"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2417","DOI":"10.1210\/endo.138.6.5164","article-title":"Preantral ovarian follicles in serum-free culture: Suppression of apoptosis after activation of the cyclic guanosine 3\u2032,5\u2032-monophosphate pathway and stimulation of growth and differentiation by follicle-stimulating hormone","volume":"138","author":"McGee","year":"1997","journal-title":"Endocrinology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1530\/REP-18-0470","article-title":"C-type natriuretic peptide enhances mouse preantral follicle growth","volume":"157","author":"Xi","year":"2019","journal-title":"Reproduction"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1097\/00004872-200018050-00010","article-title":"Oxidative stress augments secretion of endothelium-derived relaxing peptides, C-type natriuretic peptide and adrenomedullin","volume":"18","author":"Chun","year":"2000","journal-title":"J. Hypertens."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Meng, L., Zhao, K., Wang, C.C., Tao, J., Wu, Z., Teerds, K., and Zhang, S. (2021). Characterization of Long Non-Coding RNA Profiles in Porcine Granulosa Cells of Healthy and Atretic Antral Follicles: Implications for a Potential Role in Apoptosis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22052677"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1002\/ijgo.14568","article-title":"Role of C-type natriuretic peptide in polycystic ovary syndrome","volume":"161","author":"Ozay","year":"2023","journal-title":"Int. J. Gynecol. Obstet."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1042\/CS20171394","article-title":"High level of C-type natriuretic peptide induced by hyperandrogen-mediated anovulation in polycystic ovary syndrome mice","volume":"132","author":"Wang","year":"2018","journal-title":"Clin. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1002\/jcp.24682","article-title":"Brain natriuretic peptide and C-type natriuretic peptide maintain porcine oocyte meiotic arrest","volume":"230","author":"Zhang","year":"2015","journal-title":"J. Cell. Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3723","DOI":"10.1210\/en.2018-00653","article-title":"The gonadotropin-releasing hormone pulse generator","volume":"159","author":"Herbison","year":"2018","journal-title":"Endocrinology"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Constantin, S. (2017). Progress and challenges in the search for the mechanisms of pulsatile gonadotropin-releasing hormone secretion. Front. Endocrinol., 8.","DOI":"10.3389\/fendo.2017.00180"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Peng, L., He, Y., Wang, W., Chu, Y., Lin, Q., Rui, R., Li, Q., and Ju, S. (2023). PAK1 Is Involved in the Spindle Assembly during the First Meiotic Division in Porcine Oocytes. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24021123"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/20\/10046\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T14:06:07Z","timestamp":1760537167000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/20\/10046"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,15]]},"references-count":42,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["ijms262010046"],"URL":"https:\/\/doi.org\/10.3390\/ijms262010046","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,15]]}}}